Eco-friendly fabrication of Ag nanostructures using the seed extract of Pedalium murex, an ancient Indian medicinal plant: Histopathological effects on the Zika virus vector Aedes aegypti and inhibition of biofilm-forming pathogenic bacteria

The control of Zika virus mosquito vectors and well as the development of drugs in the fight against biofilm-forming microbial pathogens, are timely and important challenges in current bionanoscience. Here we focused on the eco-friendly fabrication of Ag nanostructures using the seed extract of Pedalium murex, an ancient Indian medicinal plant. Initial confirmation of Ag nanoparticles (AgNPs) production was showed by a color change from transparent to dark brown. The UV-Visible spectrum (476nm), X-ray diffraction peaks (101, 200, 220 and 311) and Fourier transform infrared spectroscopy shed light on the production of green-capped AgNPs. Morphological structure analysis using HR-TEM showed that the AgNPs were mostly hexagonal in shape with rough edges, and a size of 20-30nm. The larvicidal potential of P. murex seed extract and AgNPs fabricated using the P. murex seed extract (Pm-AgNPs) was tested on fourth instar mosquito larvae of the Zika virus vector Aedes aegypti. Maximum efficacy was achieved by Pm-AgNPs against Ae. aegypti after 24h (LC₅₀ 34.88; LC₉₀ 64.56mg/ml), if compared to the P. murex seed extract. Histopathological analyses showed severe damages to the hindgut and larval muscles in NPs-treated Ae. aegypti larvae. The sub-MIC concentrations of Pm-AgNPs exhibited significant anti-biofilm activity against Gram positive (Enterococcus faecalis, Staphylococcus aureus) and Gram negative (Shigella sonnei, Pseudomonas aeruginosa) bacterial pathogens, as showed by EPS and MTP assays. Light and CLSM microscopic studies highlighted a significant impact of P. murex seed extract and Pm-synthesized AgNPs on the surface topography and architecture of bacterial biofilm, both in Gram positive and Gram negative species. Overall, results reported here contribute to the development of reliable large-scale protocols for the green fabrication of effective mosquito larvicides and biofilm inhibitors.